miR-143-3p靶向MAPK1对人结肠癌细胞增殖,凋亡和侵袭的调节作用

The regulatory effects of miR-143-3p on proliferation, apoptosis and invasion on human colon cancer cell via targeting MAPK1

目的研究miR-143-3p靶向MAPK1与人结肠癌细胞增殖、凋亡和侵袭的关系。方法qRT-PCR检测miR-143-3p mimic转染效果和MAPK1的m RNA表达水平;双荧光素酶报告实验分析miR-143-3p与MAPK1的靶向关系;蛋白质印迹检测MAPK1的蛋白表达水平,CCK-8检测细胞增殖倍数,Hoechst染色检测细胞增殖,体外侵袭实验检测细胞侵袭,蛋白质印迹检测Ki67、VEGF、MMP-2和cleaved caspase-3的表达。结果 miR-143-3p mimic抑制人结肠癌细胞SW620中MAPK1 mRNA和蛋白的表达水平;miR-143-3p mimic与MAPK1野生型报告载体共转后,荧光素酶的活性显著降低;miR-143-3p mimic转染SW620细胞后,细胞增殖、侵袭能力显著降低,凋亡细胞数目显著增加,Ki67、VEGF和MMP-2的表达水平显著降低,cl-caspase-3的表达水平显著上升;miR-143-3p mimic能缓解MAPK1高表达对SW620细胞增殖、侵袭的诱导作用和对细胞凋亡的抑制作用。结论 miR-143-3p抑制人结肠癌细胞增殖和侵袭,诱导细胞凋亡,其作用机制与靶向抑制MAPK1有关。

Objective To investigate the relationship between miR-143-3p targeting MAPK1 and its effect on proliferation,apoptosis and invasion of human colon cancer cells.Methods qRT-PCR was used to detect the effect of transfection and the expression level of MAPK1 mRNA.Double luciferase reporter assay was used to detect the targeting relationship between microRNAs-143-3 p and MAPK1.The expression level of MAPK1 protein was detected by Western blot.The proliferation multiple was detected by CCK-8.The proliferation was detected by Hoechst staining.The cell invasion was detected by in vitro invasion test.The expression of Ki67,VEGF,MMP-2 and claspase-3 was detected by Western blot.Results miR-143-3p mimic inhibited the expression of MAPK1 mRNA and protein in SW620 cells.miR-143-3p mimic and MAPK1 wild-type reporter plasmid decreased the activity of luciferase significantly.After miR-143-3p mimic transfection,the proliferation and invasion of SW620 cells were significantly decreased,the number of apoptotic cells was significantly increased,the expression of Ki67,VEGF and MMP-2 was significantly decreased,and the expression of cl-caspase-3 was significantly increased.The miR-143-3p mimic alleviated the induction effect of high expression of MAPK1 on proliferation and invasion of SW620 cells and inhibition effect on cell apoptosis.Conclusion miR-143-3p can inhibit proliferation and invasion of human colon cancer cells and induce apoptosis through possible targeting inhibition of MAPK1.